The present invention relates to a polypeptide-polymer conjugate wherein the polymer is a graft, block, alternate, or random co-polymer coupled to the surface of the polypeptide. The invention also relates to industrial compositions and products comprising a conjugate of the invention, the use of a polypeptide-polymer conjugate of the invention for reducing the allergenicity of industrial compositions and products, and finally a method for reducing the allergenicity of polypeptides.
For both medical and industrial applications the use of polypeptides, including enzymes, are well-known in the art. As polypeptides may potentially cause an undesired immune responsexe2x80x94dependent on the way of challengexe2x80x94typically an IgG and/or IgE response, techniques for reducing it have been developed during the last three decades.
One technique is the coupling technique where a number of polymeric molecules are coupled to the polypeptide in question. When using this technique the immune system have difficulties recognizing the epitopes (on the polypeptide""s surface) responsible for the formation of antibodies, thereby reducing the immune response.
For polypeptides introduced directly into the circulatory system of the human body to give a particular physiological effect (i.e. pharmaceuticals) the typical potential immune response is an IgG and/or IgM response, while polypeptides which are inhaled through the respiratory system (i.e. industrial polypeptide) potentially may cause an IgE response (i.e. allergic response).
One of the theories explaining the reduced immune response is that the polymeric molecule(s) shield(s) epitope(s) on the surface of the polypeptide responsible for the immune response leading to antibody formation. Another theory or at least a partial factor is that the heavier the conjugate is the more reduced immune response is obtained.
Typically the polymers used for coupling to polypeptide to form conjugates are homopolymers, i.e. consisting of one repeating unit, e.g., ethylene oxide (EO), especially polyethylene glycol (PEG), or propylene oxide (PO), especially polypropylene glycol (PPG). Sacchareides, such as dextran have also been used.
U.S. Pat. No. 4,179,337 concerns non-immunogenic polypeptides, such as enzymes and peptide hormones coupled to polyethylene glycol (PEG) or polypropylene glycol (PPG).
WO 96/17929 (Novo Nordisk A/S) concerns modified polypeptide conjugates coupled to polymeric molecules, in particular polyethylene glycol (PEG).
The present invention relates to a polypeptide-polymer conjugate suitable for industrial applications and incorporation as active ingredients in industrial products.
The present inventors have found that when coupling graft, block, alternate, or random co-polymers with the general formula:
EOxPOyxe2x80x83xe2x80x83(I)
wherein x=1xe2x88x9299%, y=1xe2x88x9299%, and x+y=100% covalently to a parent polypeptide, used for industrial application, the respiratory allergenicity is reduced when compared to the parent enzyme and even when compared to a corresponding conjugate coupled with PEG or other homopolymers.
In a second aspect the invention relates to compositions for use in industrial products comprising a conjugate of the invention.
In a third aspect the invention relates to a method for reducing the allergenicity of polypeptides.
Polypeptides used for industrial applications often have an enzymatic and/or anti-microbial activity. Industrial polypeptides are (in contrast to pharmaceutical polypeptides) not intended to be introduced into the circulatory system of the body.
Therefore, it is not very likely that industrial polypeptides, such as enzymes, used as active ingredients in industrial compositions and/or products (defined below), such as detergents, such as laundry and dish washing detergens, food or feed additives, including additives for bread making, composition for treating textiles, and personal care products, including cosmetics, come into direct contact with the circulatory system of the body of humans or animals, as such polypeptides (or products comprising such polypeptides) are not injected (or the like) into the bloodstream.
Thus, in the case of the industrial polypeptide the potential risk is respiratory allergy (i.e. IgE response) as a consequence of inhalation of polypeptides through the respiratory passage.
In the context of the present invention xe2x80x9cindustrial polypeptidesxe2x80x9d are defined as polypeptides, including peptides, proteins and/or enzymes, which are not intended to be introduced into the circulatory system of the body of humans and/or animals.
Examples of such polypeptide include polypeptides with enzymatic activity as defined below.
The invention also relates to industrial compositions and products having reduced respiratory allergenicity.
Furthermore, the invention relates to the use of a polypeptide-polymer conjugate of the invention for reducing the respiratory allergenicity of industrial composition and products.
Finally the invention relates to a method for reducing the allergenicity of polypeptides, in particular enzymes, by coupling one or more a graft, block, alternate, or random co-polymer to a parent unmodified polypeptide.